Somatostatin-producing cerebral cortical cell cultures were grown in either high- (33 mM) or low-glucose (5 mM) medium and then exposed to short repetitive changes of high- or low-glucose Krebs-Ringer's bicarbonate buffer. Equivalent amounts of somatostatin were released in the high-to-high, the low-to-low, and the low-to-high paradigms. The high-to-low experiment produced a rapid rise in somatostatin release, followed by a decline. Cultures exposed to 2-deoxyglucose after high-glucose medium also released much greater amounts of immunoreactive somatostatin. Separate sets of cultures were grown in high- or low-glucose medium for up to 19 days. Cultures grown in high-glucose medium generally contained more somatostatin intracellularly than did those maintained in low glucose, although somatostatin in the medium was only different at day 19. These results identify extracellular glucose as an important determinant of cortical somatostatin production.